Freezing and thawing processes may damage many cells and nuclei, potentially leading to the loss of nuclear components or contamination of the nuclei with cytoplasmic or other cellular components.
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NUC101
Zellkernisolierung - Kit: Nuclei EZ Prep
sufficient for 25 nuclei preparations (~1-10×107 cells/preparation)
Synonym(e):
Rapid mammalian nuclei isolation kit
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1 KIT
400,00 €
400,00 €
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1 KIT
400,00 €
About This Item
UNSPSC-Code:
12352207
NACRES:
NA.32
400,00 €
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Empfohlene Produkte
Verwendung
sufficient for 25 nuclei preparations (~1-10×107 cells/preparation)
Qualitätsniveau
Haltbarkeit
1 yr at 2‑8 °C
Verpackung
pkg of 1 kit
Anwendung(en)
cell analysis
Fremdaktivität
nuclease and protease, free
Versandbedingung
wet ice
Lagertemp.
2-8°C
Allgemeine Beschreibung
Das Nuclei EZ Prep-Kit wurde speziell für die rasche Isolierung der Zellkerne aus Säugetierzellen entwickelt. Das Kit liefert einen hohen Ertrag an Zellkernen aus häufig verwendeten Zelltypen.
Anwendung
Geeignete Quelle für Nuklearkomponenten zur Produktion von Zellkernen für In-vitro-Apoptose-Assays und für funktionelle Studien.
Biochem./physiol. Wirkung
Das Protokoll liefert einen hohen Ertrag an häufig verwendeten Säugetierzellkernen, sowohl von adhärenten (z. B. HEK293 und COS7) als auch nicht adhärenten (z. B. Jurkat und HFN7.1) Zelllinien und peripheren mononuklearen Blutzellen (PBMC). Die Präparationen eignen sich für eine Vielzahl von Zellbiologie-Anwendungen, z. B. als Quelle für Nuklearkomponenten wie Chromatin, genomische DNA, Histone und nukleäre RNA/RNP, die Produktion von Zellkernen für In-vitro-Apoptose-Assays und funktionelle Studien, wie die Untersuchung des Transkriptionsstatus von Zellen.
Sonstige Hinweise
Alle Verfahren sollten auf Eis oder bei 2 - 8 °C durchgeführt werden.
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Lagerklassenschlüssel
10 - Combustible liquids
Flammpunkt (°F)
Not applicable
Flammpunkt (°C)
Not applicable
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Analysis of nuclear RNA, Chapter 14.
Robert E. Farrell, Jr., ed.
RNA Methodologies: A Laboratory Guide for Isolation and Characterization, 235-263 (1993)
Christina N Vallianatos et al.
Communications biology, 3(1), 278-278 (2020-06-03)
Histone H3 lysine 4 methylation (H3K4me) is extensively regulated by numerous writer and eraser enzymes in mammals. Nine H3K4me enzymes are associated with neurodevelopmental disorders to date, indicating their important roles in the brain. However, interplay among H3K4me enzymes during
Yu-Yin Shih et al.
PloS one, 6(10), e26236-e26236 (2011-10-25)
Retinoic acid (RA) has been approved for the differentiation therapy of neuroblastoma (NB). Previous work revealed a correlation between glucose-regulated protein 75 (GRP75) and the RA-elicited neuronal differentiation of NB cells. The present study further demonstrated that GRP75 translocates into
Buqing Ye et al.
Nature communications, 8(1), 1518-1518 (2017-11-16)
Lymphoid lineage commitment is an important process in haematopoiesis, which forms the immune system to protect the host from pathogen invasion. However, how multipotent progenitors (MPP) switch into common lymphoid progenitors (CLP) or common myeloid progenitors (CMP) during this process
Naomi Habib et al.
Nature methods, 14(10), 955-958 (2017-08-29)
Single-nucleus RNA sequencing (sNuc-seq) profiles RNA from tissues that are preserved or cannot be dissociated, but it does not provide high throughput. Here, we develop DroNc-seq: massively parallel sNuc-seq with droplet technology. We profile 39,111 nuclei from mouse and human
Artikel
Centrifugation separates organelles based on size, shape, and density, facilitating subcellular fractionation across various samples.
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Can this buffer be used for frozen human lung tissue?
1 Antwort-
Hilfreich?
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Can this buffer be used with ipscs and can the incubation be conducted in a thermocyler at 2-4C instead of on ice
1 Antwort-
Yes, this kit has been referenced to work with induced pluripotent stem cells (iPSCs). Although not tested, using a thermocycler set to 2-4°C will effectively maintain the required conditions as a viable alternative to using ice for the incubation steps during the nuclei isolation process. Please see the link below to review the publication referencing use with iPSCs.
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6992778/Hilfreich?
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What is the pH of the EZ-prep lysis buffer
1 Antwort-
The complete formulation for the Lysis solution is given in the following reference. The lysis buffer in the kit is formulated the same, except without the Triton X-100 and the DTT.
“Analysis of nuclear RNA”, Chapter 19, in RNA Methodologies: A Laboratory Guide for Isolation and Characterization, Robert E. Farrell, Jr., Academic Press, San Diego, p. 406-437 (1998)Hilfreich?
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Is the Nuclei isolation Kit Nuclei EZ Prep designed for DNA or protein isolation?
1 Antwort-
The Nuclei isolation Kit Nuclei EZ Prep is designed for the isolation of intact nuclei containing nucleic acid and protein factors.
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What is the formulation of the storage solution of NUC101?
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The storage solution for NUC101 (Product No. S8933) contains 2 mM MgCl2, 0.1 mM Na2EDTA, and no detergent in a Tris buffer (pH 8.0) with 30% glycerol.
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Can this kit be used with vertebrate cells which are not mammalian (such a avian cells)?
1 Antwort-
Product NUC101 has been validated for use with mammalian samples. However, it has been used in many different species by independent laboratories. One study used Nuc201 with zebrafinch samples:
https://www.nature.com/articles/s41467-021-22918-2Hilfreich?
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For the upcoming important phase of our project, we need the Nuclei EZ Prep Nuclei Isolation Kit. Do you have alternatives to this product in your range, which we can use for our Nuclei Isolation? Or are there any remaining stocks that we can use.
1 Antwort-
The NUC201 Nuclei Isolation Kit is an available alternative for this product. The NUC201 method includes centrifugation in a sucrose solution which offers additional protection to the nuclei, but may result in a lower yield when compared to the NUC101. Please see the link below to review the product datasheet:
https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/product/documents/210/075/nuc201bul.pdfFor additional questions regarding this product option, we kindly ask you to navigate to the link https://www.sigmaaldrich.com/techservice, click on "Product Technical Inquires" under the Products Section with all the required information so that a member of our team can reach out to you to assist further. Thank you.
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What is the Department of Transportation shipping information for this product?
1 Antwort-
Transportation information can be found in Section 14 of the product's (M)SDS.To access the shipping information for this material, use the link on the product detail page for the product.
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Should I use trypsin to harvest my adherent cells before using Product NUC101, Nuclei Isolation Kit?
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In general, this is not advised. We have never tried detaching the cells first with trypsin. We would not recommend it, because residual trypsin could clip nuclear proteins and cause problems in downstream applications.
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Can Product NUC101, Nuclei Isolation Kit, be used to capture soluble nuclear proteins?
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The kit was originally designed to produce nuclei that are competent for transcription run-on/run-off assays to measure transcription from pre-engaged polymerase complexes on the nuclear DNA. It was not designed for the separation or isolation of soluble nuclear proteins. The researcher might be advised to try the NXTRACT nuclear extraction kit, which allows for the isolation of "intact" nuclei from other cellular components before the lysis of the nuclear membrane.
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